pFC83 [CMCC 3128] (ATCC® 67422)

Organism: Thermus aquaticus Brock and Freeze  /  Clone Type: Clone  /  Depositors: Cetus Corp., A Belt, Cetus Corp.

Permits and Restrictions

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Designations pFC83 [CMCC 3128]
GenBank Number

J04639

Species Thermus aquaticus Brock and Freeze
Depositors Cetus Corp., A Belt, Cetus Corp.
Vector
Construct size (kb): 7.699999809265137
Insert
DNA: genomic
Insert lengths(kb): 4.5
Gene product: Taq DNA polymerase [polI]
Insert Size (kb) 4.5
Media Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Biosafety Level 1
Shipping Information

Shipped:  Freeze dried phagemid in E. coli

Comments
Restriction digests of the clone give the following sizes (kb): HindIII--4.6, 3.4; BamHI--5.1, 3.0; BglII--8.0.
Higher yields of the plasmid DNA can be obtained when the culture is grown at 30 C.
The insert contains a BamHI and a BglII site.
The sequence of the BglII/BamHI fragment is given as nucleotides -112 to +614 in the published sequence. This encodes the amino terminal portion of the protein.
A full-length Taq polymerase gene can be generated by ligating the HindIII-Asp718 fragment from pFC85 (ATCC 67421) to a 750 bp BglII-HindIII fragment from pFC83 (ATCC 67422).
References

Lawyer FC, et al. Isolation, characterization, and expression in Escherichia coli of the DNA polymerase gene from Thermus aquaticus. J. Biol. Chem. 264: 6427-6437, 1989. PubMed: 2649500

Gelfand DH, et al. Purified thermostable enzyme. US Patent 4,889,818 dated Dec 26 1989

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